The 96-well microplate and related versions thereof, such as 384-well microplates, continue to play a critical role in the field of analytical assays. Indeed, such microplates are ubiquitously employed in research laboratories, analytical laboratories, and medical diagnostic laboratories throughout the world. For many years, there has been a growing demand for methods that enable investigators and laboratories to perform multiplex assays using these standard microplate formats.
A multiplex assay is a type of procedure that simultaneously—in a single assay—measures, detects and/or analyzes multiple analytes. Multiplex assays have been used in order to detect or quantify various biomolecules in a particular sample, such as mRNAs, proteins, antibodies, and other biomolecules. Multiplex assay formats are often beneficial, insofar as such formats can provide a significant reduction in assay costs, on a cost-per-analyte basis. In addition, such formats significantly increase the amount (and often types) of information that can be extracted from each sample, particularly on a per-sample-volume basis.
Despite the significant utility of multiplex assay formats, present platforms do not allow for the dispensing of a specific secondary binding agent (i.e., the detection agent) to each of a plurality of immobilized targets, in order to reduce cross-reactivity (which leads to false positive results). This drastically limits the types of assays that may be combined in a multiplex fashion (and, more particularly, the combination of analytes that may be measured or detected in a single assay format). In addition, current platforms do not allow individual assay conditions, e.g., sample dilutions, buffer types, incubation times, etc., to be optimized. Accordingly, a continuing need exists for new and improved multiplex binding assay assemblies and methods of use thereof.